Balasubramanian N, Juliet G Annie, Srikalaivani P, Lalithakumari D
Center for Advanced Studies in Botany, University of Madras, Chennai, India.
Can J Microbiol. 2003 Apr;49(4):263-8. doi: 10.1139/w03-034.
A protocol for isolating and regenerating protoplasts from Trichothecium roseum has been described. Protoplasts from T. roseum were isolated using (i) a lytic enzyme combination composed of Novozym 234, chitinase, cellulase, and pectinase at a 5-mg/mL concentration and (ii) 0.6 M KCl as an osmotic stabilizer. A maximum number of 28 x 10(4) protoplasts/mL were obtained at pH 5.5. Experiments on the regeneration and reversion of protoplasts revealed a maximum regeneration (60.8%) in complete medium (potato dextrose--yeast extract agar) amended with 0.6 M KCl. The regenerated protoplasts were similar to the original parent strain in morphology, pigmentation, growth, and sporulation.
已描述了一种从粉红单端孢霉中分离和再生原生质体的方案。使用（i）由浓度为5mg/mL的诺维信234、几丁质酶、纤维素酶和果胶酶组成的裂解酶组合，以及（ii）0.6M KCl作为渗透稳定剂，从粉红单端孢霉中分离原生质体。在pH 5.5时，获得的原生质体数量最多为28×10⁴个/mL。原生质体再生和回复的实验表明，在添加了0.6M KCl的完全培养基（马铃薯葡萄糖 - 酵母提取物琼脂）中，最大再生率为60.8%。再生的原生质体在形态、色素沉着、生长和孢子形成方面与原始亲本菌株相似。
